Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-M6PR (cation dependent) antibody [EPR7691] (ab134153)

Reviews (2)Q&A (1)References (3)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR7691] to M6PR (cation dependent)
  • Suitable for: WB, Flow Cyt, ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-M6PR (cation dependent) antibody [EPR7691]
    See all M6PR (cation dependent) primary antibodies

  • Description

    Rabbit monoclonal [EPR7691] to M6PR (cation dependent)

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, Flow Cyt, ICC/IFmore details
    Unsuitable for: IHC-P

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide corresponding to Human M6PR (cation dependent) aa 250 to the C-terminus (C terminal).
    Database link: P20645

  • Positive control

    • WB: HAP1, A549, and Human uterus lysates ICC/IF: HeLa cell lysate Flow Cyt: A549 cells

  • General notes

     

     

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

Applications

Our Abpromise guarantee covers the use of ab134153 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/1000 - 1/10000. Predicted molecular weight: 31 kDa.
Flow Cyt 1/80.
ICC/IF 1/50 - 1/100.
  • Application notes
    Is unsuitable for IHC-P.

    • Target

    • Function

      Transport of phosphorylated lysosomal enzymes from the Golgi complex and the cell surface to lysosomes. Lysosomal enzymes bearing phosphomannosyl residues bind specifically to mannose-6-phosphate receptors in the Golgi apparatus and the resulting receptor-ligand complex is transported to an acidic prelyosomal compartment where the low pH mediates the dissociation of the complex.

    • Domain

      The extracellular domain is homologous to the repeating units (of approximately 147 AA) of the cation-independent mannose 6-phosphate receptor.

    • Cellular localization

      Lysosome membrane.
    • Information by UniProt

    • Database links

      see all

    • Alternative names

      • 46 kDa mannose 6 phosphate receptor antibody
      • 46 kDa mannose 6-phosphate receptor antibody
      • Cation dependent mannose 6 phosphate receptor antibody
      • Cation dependent mannose 6 phosphate receptor precursor antibody
      • Cation-dependent mannose-6-phosphate receptor antibody
      • CD Man 6 P receptor antibody
      • CD Man-6-P receptor antibody
      • CD MPR antibody
      • CD-M6PR antibody
      • CD-MPR antibody
      • FLJ32994 antibody
      • M6pr antibody
      • M6PR protein antibody
      • Man6PR antibody
      • Mannose 6 phosphate receptor (cation dependent) antibody
      • Mannose 6 phosphate receptor antibody
      • MPR 46 antibody
      • MPR46 antibody
      • MPRD antibody
      • MPRD_HUMAN antibody
      • Mr 46,000 Man6PR antibody
      • Small mannose 6 phosphate receptor antibody
      • SMPR antibody
      see all

    Images

    • Western blot - Anti-M6PR (cation dependent) antibody [EPR7691] (ab134153)
      Western blot - Anti-M6PR (cation dependent) antibody [EPR7691] (ab134153)

      Lane 1: Wild type HAP1 whole cell lysate (20 µg)
      Lane 2: Empty
      Lane 3: M6PR knockout HAP1 whole cell lysate (20 µg)
      Lane 4: A549 whole cell lysate (20 µg)

      Lanes 1 - 4: Merged signal (red and green). Green - unpurified ab134153 observed at 46 kDa. Red - loading control, ab18058, observed at 130 kDa.

      ab134153 was shown to specifically react with M6PR when M6PR knockout samples were used. Wild-type and M6PR knockout samples were subjected to SDS-PAGE. ab134153 and ab18058 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

    • Immunocytochemistry/ Immunofluorescence - Anti-M6PR (cation dependent) antibody [EPR7691] (ab134153)
      Immunocytochemistry/ Immunofluorescence - Anti-M6PR (cation dependent) antibody [EPR7691] (ab134153)
      Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling M6PR (cation dependent) with Purified ab134153 at 1:100 dilution (8.6 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor®594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor®488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
    • Western blot - Anti-M6PR (cation dependent) antibody [EPR7691] (ab134153)
      Western blot - Anti-M6PR (cation dependent) antibody [EPR7691] (ab134153)
      All lanes : Anti-M6PR (cation dependent) antibody [EPR7691] (ab134153) at 0.8 µg/ml (purified)

      Lane 1 : A549 (Human lung carcinoma epithelial cell) whole cell lysates
      Lane 2 : Mouse kidney lysates
      Lane 3 : Rat kidney lysates
      Lane 4 : Rat spleen lysates

      Lysates/proteins at 15 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

      Predicted band size: 31 kDa



      Blocking and diluting buffer: 5% NFDM/TBST.

    • Flow Cytometry - Anti-M6PR (cation dependent) antibody [EPR7691] (ab134153)
      Flow Cytometry - Anti-M6PR (cation dependent) antibody [EPR7691] (ab134153)
      Flow Cytometry analysis of A549 (Human lung carcinoma epithelial cell) cells labeling M6PR (cation dependent) with purified ab134153 at 1:80 dilution (10 µg/ml) (red). Cells were fixed with 4% Paraformaldehyde. A Goat anti rabbit IgG (Alexa Fluor® 488) secondary antibody was used at 1:2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
    • Western blot - Anti-M6PR (cation dependent) antibody [EPR7691] (ab134153)
      Western blot - Anti-M6PR (cation dependent) antibody [EPR7691] (ab134153)
      All lanes : Anti-M6PR (cation dependent) antibody [EPR7691] (ab134153) at 1/1000 dilution (Unpurified)

      Lane 1 : A549 lysates
      Lane 2 : Human uterus lysates

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution

      Predicted band size: 31 kDa

    • OI-RD Scanning - Anti-M6PR (cation dependent) antibody [EPR7691] (ab134153)
      OI-RD Scanning - Anti-M6PR (cation dependent) antibody [EPR7691] (ab134153)
      Equilibrium disassociation constant (KD)
      Learn more about KD

      Click here to learn more about KD
    • Anti-M6PR (cation dependent) antibody [EPR7691] (ab134153)
      Anti-M6PR (cation dependent) antibody [EPR7691] (ab134153)

    References (3)

    Publishing research using ab134153? Please let us know so that we can cite the reference in this datasheet.

    ab134153 has been referenced in 3 publications.

    • Cui L  et al. The Lysosomal Membrane Protein Lamp2 Alleviates Lysosomal Cell Death by Promoting Autophagic Flux in Ischemic Cardiomyocytes. Front Cell Dev Biol 8:31 (2020). PubMed: 32117965
    • Han J  et al. HSP90 inhibition targets autophagy and induces a CASP9-dependent resistance mechanism in NSCLC. Autophagy 14:958-971 (2018). PubMed: 29561705
    • Di Stefano AL  et al. Detection, Characterization, and Inhibition of FGFR-TACC Fusions in IDH Wild-type Glioma. Clin Cancer Res 21:3307-17 (2015). PubMed: 25609060

    Customer reviews and Q&As

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    Question

    Literature shows that band of glycosylated protein should be around 45 kDa, not 31 kDa.

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    Answer

    Thank you for your call today and for letting us know about the trouble with ab76276.

    As we discussed, I'm sending a free of charge vial of ab134153 on the order ***, which I expect to arrive by July 9. Please keep me updated about the results with this antibody.

    I look forward to hearing from you. If there is anything else that I can dofor you in the meantime, please let me know and I'll be happy to help.

    Have a nice day.

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